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Biomedical subjects

S Minucci

Publications and source records attributed to S Minucci.

At least 91 records · Page 5Linked to original sources

The effects of gonadectomy and testosterone treatment on the Harderian gland of the green frog, Rana esculenta.

The effects of gonadectomy and testosterone treatment on the fine structure of the Harderian gland in male and female green frogs were investigated in different periods of the year. Gonadectomy, carried out when the glands are in the lowest secretory phase (September), causes degenerative changes consisting of a reduction of the rough endoplasmic reticulum, the appearance of autolysosomes, and an increase of nuclear heterochromatin. These effects can be prevented by testosterone treatment. No castration effects are found during the recovery (November) and enhancement (April-May) phases of secretory activity. The results suggest that the frog Harderian gland's sensitivity to testosterone changes during the annual cycle. The androgen dependence of the Harderian gland is correlated with the presence of androgen receptors in both male and female frogs.

Animals↗

The orbital glands of the terrapin Pseudemys scripta in response to osmotic stress: a light and electron microscope study.

The histochemical and ultrastructural features of both orbital glands--the anterior lacrimal and harderian glands--were compared in the terrapin Pseudemys scripta following adaptation either to distilled water or to seawater. Seawater adaptation activated the harderian gland and caused a major structural reorganisation. Type I cells increased in number, whereas type II cells became smaller and less numerous. The most striking change was shown by the type IV cells which display the features of a salt-secreting cell. This cell type increased in number and changed from a unicellular form to multicellular complexes. Transfer from fresh water to distilled water caused a decrease in alcianophilia in type I and type III glandular cells and degenerative phenomena were often seen in type I and type IV cells. The anterior lacrimal gland showed only minor changes either in distilled or seawater adapted terrapins. Transfer to distilled water also caused degenerative phenomena in the lacrimal gland. These results establish for the first time the involvement of the harderian gland of a terrapin in osmoregulation.

Adaptation, Physiological↗

Intratesticular control of spermatogenesis in the frog, Rana esculenta.

Adult intact and hypophysectomized (PDX) frogs, Rana esculenta, were treated with a gonadotropin releasing hormone agonist (GnRHA, HOE 766) and/or cyproterone acetate (CPA), the antiandrogen, in order to investigate the regulation of primary spermatogonial (I SPG) multiplication in vertebrates. Treatment with GnRHA (injections containing 900 ng administered for 12 days on alternate days) caused a significant increase of the mitotic index (MI) of I SPG in PDX animals and a further MI increase of SPG was observed when 0.66 mg CPA was given concomitantly with GnRHA. The treatment with 0.66 mg CPA in combination with GnRHA also increased secondary spermatocyte (II SPC) appearance. Moreover, number of nests containing spermatids (SPT) decreased as CPA, in combination with GnRHA, was administered in increasing doses (0.33 and 0.66 mg/injection). Intact animals treated with CPA (0.66 mg/injection) showed a time-dependent I SPG multiplication increase which reached highest values after 28 days. Secondary SPC also proliferated until day 28; meanwhile the number of nests containing SPT decreased. Neither testosterone nor R5020 (a progestin which is not converted to androgens) modified the basal and GnRHA-induced spermatogonial proliferation. These results confirm that in the frog, Rana esculenta, spermatid formation is impaired by CPA treatment and that I SPG multiplication is enhanced by a direct effect of GnRHA; moreover, we suggest that the absence of spermatids constitutes a signal promoting spermatogonial proliferation.

Animals↗

Immunocytochemical identification of some regulatory peptides (gastrin, gastrin-releasing peptide, neurotensin and vasoactive intestinal polypeptide) in the Harderian gland of the green frog, Rana esculenta.

The presence and distribution of gastrin-, gastrin-releasing peptide-, neurotensin- and vasoactive intestinal polypeptide-like immunoreactivity in the Harderian gland of Rana esculenta were studied at different times of the annual cycle. Gastrin-releasing peptide, neurotensin and vasoactive intestinal polypeptide-like substances were found either in the glandular cells, or in the nerve fibers surrounding the glandular acini. Gastrin-like immunoreactivity was confined to the glandular cells. The immunoreactivity varied during the annual cycle, with the greatest concentration being noted during the recovery phase of glandular secretory activity.

Animals↗

Ultrastructural investigation of the corpora atretica of the electric ray, Torpedo marmorata.

Follicular atresia was studied in the ovary of the electric ray, Torpedo marmorata, by light and electron microscopy. The course of atresia may be divided into four stages. The first two comprise the dissolution of the oocyte and its phagocytosis by the small cells of the granulosa epithelium. The third stage consists of the transformation of the granulosa epithelium into an active glandular structure and is accompanied by the development of a smooth endoplasmic reticulum. The fourth stage is marked by sclerosis and pigmentary degeneration of the atretic follicle. Together these observations suggest an endocrine steroidogenic role for the corpora atretica (preovulatory corpora lutea) in T. marmorata.

Animals↗

Testosterone induction of poly(A)(+)-RNA synthesis and [35S]methionine incorporation into proteins of Rana esculenta Harderian gland.

The role of androgens in the cyclic secretory activity of the Rana esculenta Harderian gland (HG) was studied. Total RNA showed a dramatic increase in October and May when the nuclear androgen receptors peak. During the resumption of the secretory activity a gradual increase of poly(A)(+)-RNA was detected; during the enhancement phase (May) a peak of the poly(A)(+)-RNA fraction was found. In in vitro experiments testosterone increased the incorporation of [3H]uridine into the poly(A)(+)-RNA fraction and also that of [35S]methionine into a newly synthesized protein fraction (100 kDa). The latter effect is prevented by the exposure of the cells to the antiandrogen, cyproterone acetate (CPA). These findings reveal that, besides hamsters, the HG is a target for androgens in the frog.

Animals↗

Effect of castration and testosterone therapy on harderian gland protein patterns of the golden hamster (Mesocricetus auratus).

1. Sodium dodecyl sulphate 7-12% gradient polyacrylamide gel electrophoresis of male and female hamster Harderian gland whole homogenate shows a clear-cut sexual dimorphism, which consists of the presence of two male-specific glycoproteins (168 and 116 kDa) and two specific female proteins (210 and 190 kDa). 2. In the male, castration causes a significant decrease in the concentration of the two glycoprotein fractions. 3. Replacement therapy with testosterone propionate (T) restores the intact male pattern.

Animals↗

Resumption of testicular activity in Gobius paganellus after administration of ethane 1,2-dimethane sulfonate (EDS).

1. The effect of a single injection of ethane-1,2-dimethane sulfonate (EDS) was studied in the teleost fish, Gobius paganellus in two different periods of the year. 2. During June EDS did not induce any change, while during December the drug was highly effective in promoting testicular activity. 3. Nucleus/cytoplasm ratio of interstitial cells strongly decreased concomitantly with the detection of high testicular androgen levels. 4. The germinal compartment was well developed showing the appearance of all spermatogenic stages and the cavity of lobular compartments filled of spermatozoa. 5. Our data are the first evidence of a stimulatory activity of EDS on testes of a vertebrate species.

Androgens↗

Mallory stain may indicate differential rates of RNA synthesis: I. A seasonal cycle in the harderian gland of the green frog (Rana esculenta).

When Mallory's trichrome stain is used, acinar nuclei of the Harderian gland of Rana esculenta display different affinities for the dye. Some of the orangiophilic nuclei show affinity for aniline blue (blue nuclei). In the Harderian gland of Rana esculenta their number and the intensity of staining with aniline blue may vary during the year. The affinity for aniline blue disappears following digestion of paraffin sections with RNAase, but not with DNAase or trypsin. Furthermore, in vitro incubation with [5, 6-3H]-Uridine shows a selective incorporation by the majority of blue nuclei. Therefore, the affinity for aniline blue is likely due to increased RNA synthesis. The increment of nuclear RNA shown by these methods is supported by the quantitative determination of total RNAs during the resumption (October) and enhancement (May) of secretory activity, when the percentage of blue nuclei of the acinar cells is at its highest levels of the year. The affinity of RNA-rich nuclei for aniline blue, while others are strictly orangiophil, is discussed on the basis of molecular structure of the dyes used in the staining mixture. Mallory's trichrome stain appears to be an useful tool for detecting changes in cell nuclear status.

Animals↗

Mallory stain may indicate differential rates of RNA synthesis: II. Comparative observations in vertebrate nuclei.

The differential staining of nuclei by the use of the Mallory trichrome method was investigated in a variety of tissues of representative vertebrates. By this method nuclei stained orange or blue; erythrocyte nuclei stained red. Since the higher affinity for aniline blue is due to an increased RNA synthesis, it was possible to reveal not only the changing metabolic status of a cell type, as shown for instance in the liver parenchyma and other glandular tissues, and nervous tissue, but also in different cell populations in the same tissue, such as the spleen.

Amphibians↗

The orbital glands of the chelonians Pseudemys scripta and Testudo graeca: comparative histological, histochemical and ultrastructural investigations.

The orbital glands of the chelonians Pseudemys scripta and Testudo graeca were investigated at the histological, histochemical and ultrastructural levels. Four acinar cell types were seen in the harderian gland of P. scripta on the basis of histochemical reactions and ultrastructure. Secretory granules were of 2 types, one showing moderate electron density with an electronlucent core, the other being smaller and more osmiophilic with an electron-dense core. In the harderian gland of T. graeca only 2 glandular cell types were found; one type contained secretory granules with a dense core surrounded by a wide zone of lower density. Acinar cells of the anterior lacrimal gland in both species were of 2 types, one being of mucous type. In the harderian gland and in the lacrimal gland of both species, one cell type appeared not to be involved in the secretion of organic material. These cells contained numerous tightly packed mitochondria among which were abundant clumps of glycogen; the cell membrane was specialised at both edges. This cell type was similar ultrastructurally to the 'salt cells' described in the salt-secreting glands of various marine vertebrates, i.e. of the cells involved in transport processes. These combined histological, histochemical and ultrastructural studies have allowed us to distinguish orbital glands. In the past, the harderian and lacrimal glands in chelonians have often been mistaken for one another.

Anatomy, Comparative↗

Proteolytic activity of the purified hormone-binding subunit in the estrogen receptor.

The hormone-binding subunit of the calf uterus estradiol receptor was purified as a hormone-free molecule. Immunoaffinity chromatography with a specific monoclonal antibody was used as the final step. The purified subunit was specifically labeled by radioactive diisopropyl fluorophosphate. The diisopropyl fluorophosphate-labeled amino acid was serine. The purified receptor was able to release the fluorogenic or chromogenic group from synthetic peptides containing phenylalanine at the carboxyl terminus. This occurred only in the presence of estradiol and was hampered by aprotinin and diisopropyl fluorophosphate. Estradiol-dependent hydrolytic activity was also found in the eluate from gel slices after SDS/PAGE of purified receptor. This activity comigrated with the renaturable estradiol-binding activity. The estradiol antagonists 4-hydroxytamoxifen and ICI 164,384 as well as other steroid hormones were unable to activate this hydrolytic activity.

Amino Acid Sequence↗

Androgen receptor in the Harderian gland of Rana esculenta.

An androgen receptor has been identified in the cytosolic and nuclear extracts of the Harderian gland of the frog, Rana esculenta. A single class of high-affinity binding sites was found: Kd = 1.9 +/- 1.3 (S.D.) nmol/l (n = 26) for the cytosolic extract and Kd = 0.9 +/- 0.8 nmol/l (n = 15) for the nuclear extract. The presence of binding activity in both nuclear and cytosolic extracts and the low rate of ligand-receptor dissociation are characteristics that distinguish this receptor from a steroid-binding protein. The Kd did not show any sex difference and did not exhibit any secretory activity-related change. Binding in both cytosolic and nuclear extracts was specific for androgens (testosterone = 5 alpha-dihydrotestosterone); oestradiol-17 beta showed a 30% cross-reaction; moreover, specific binding of [3H]oestradiol-17 beta was not detectable. The binding capacity of the Harderian gland increased progressively in both fractions from October to December, reaching a peak in May, and decreased suddenly during July to August. The lack of any morphological sex-related difference in the Harderian gland of the green frog might be accounted for by the high amount of circulating androgens as well as a similar concentration of androgen receptor in both sexes.

Animals↗

Number of mast cells in the harderian gland of the green frog, Rana esculenta: the annual cycle and its relation to environmental and hormonal factors.

The Harderian gland of the green frog contains mast cells. Their number shows annual variations, being more numerous in the winter months. The increase of mast cell number (MCN) is matched by a marked degranulation. No sex differences are found throughout the year. Manipulations of the photoperiod and temperature, either in winter or in summer, suggest that only the latter is responsible for the annual variations. Exposure to higher temperatures causes a decrease in the MCN in the winter frogs, while exposure of the summer frogs to low temperatures provokes the opposite effect. The pituitary gland also influences MCN. Hypophysectomy causes a decrease of MCN, with a return to normal following replacement therapy with homologous pars distalis homogenate. Among pituitary hormones, only ACTH mimics the effect of pars distalis homogenate. However, a possible link seems to exist between environmental (temperature) and hormonal (pituitary) factors, since hypophysectomy prevents the increase of MCN in the summer frogs exposed to low temperatures.

Animals↗

An aprotinin binding site localized in the hormone binding domain of the estrogen receptor from calf uterus.

It has been proposed that the estrogen receptor bears proteolytic activity responsible for its own transformation. This activity was inhibited by aprotinin. Incubation of transformed ER with aprotinin modified the proteolytic digestion of the hormone binding subunit by proteinase K. The smallest hormone-binding fragment of the ER, obtained by tryptic digestion, was still able to bind to aprotinin. These results suggest that aprotinin interacts with ER and the hormone-binding domain of ER is endowed with a specific aprotinin-binding site.

Animals↗

Harderian gland and the lacrimal gland of the lizard Podarcis s. sicula: histology, histochemistry, and ultrastructure.

Histology, histochemistry, and ultrastructure of the Harderian gland and lacrimal gland of the lizard Podarcis s. sicula were investigated. The Harderian gland, located at the medial corner of the orbit, can be divided into three zones showing different tinctorial features either with Mallory or hematoxylineosin stains. The glandular cells of the acinar medial zone secrete predominantly acidic sulphated mucosubstances. The acinar cells of the intermediate zone contain secretory granules that show a weak reaction to the histochemical tests for mucosubstances. The lateral zone has a tubulo-acinar type of structure and tests strongly for proteins, whereas Alcian-PAS staining is very weak. The lacrimal gland is smaller than the Harderian gland and lies in the region of the posterior commissure of the eyelids. it shows the same histological and histochemical characteristics of the medial zone of the Harderian gland, i.e., it is mucous secreting. At the ultrastructural level the zonation is well defined, especially when the secretory granules are examined. Granules of the mucoid type are found in the lacrimal gland and the medial zone of the Harderian gland. The secretory granules of the lateral part of the Harderian gland show a composite structure never described before. Therefore, they have been called "special secretory granules." Each of these granules is composed of three sharply separated components. It is not known whether the three components correspond to different secretions. Histochemical tests suggest that they are of the serous type. Both mucous and serous granules are secreted by the same glandular cells of the intermediate zone of the Harderian gland. The two types of granules usually occupy different cell compartments. The mechanism of secretion appears either merocrine or apocrine in both the Harderian gland and the lacrimal gland.

Animals↗

Morphological and hormonal changes in the frog, Rana esculenta, testis after administration of ethane dimethane sulfonate.

Apart from mice, in rodents ethane dimethane sulfonate (EDS) selectively destroys Leydig cells. This has been indicated as a new method for the study of seminiferous interstitial compartment interaction. No information on the possible destruction and repopulation of Leydig cells exists in lower vertebrates. This study deals with EDS effects in the frog, Rana esculenta. Animals received a single intraperitonial dose (100 mg/kg body wt) and were sacrificed at 0, 12, and 24 hr and 3, 4, 7, 14, and 28 days postinjection. Androgens (testosterone + DHT) were measured in plasma and right testes. Moreover, left testes were fixed and examined for histological observation. Plasma androgen levels were extremely low on Day 4 after EDS treatment and remained unchanged thereafter. In testes, androgen levels decreased on Day 4 but increased to control levels on Day 14. Leydig cells were damaged within 3 days post-treatment and were completely destroyed on Days 4 and 5. Germinal compartment damage appeared only where the adjacent interstitial tissue presented complete destruction. Pale primary spermatogonia (stem cells) were always present. Testes restored to normal on Day 14 and spermatogenesis resumed to the regenerating interstitial tissue. These results show that regenerating testes in R. esculenta retain androgens and that interstitial-germinal compartment communications may have a role in maintaining spermatogenesis.

Androgens↗